Identification of Endogenous Genes for Normalizing Titer Variation of Citrus Tristeza Virus in Aphids at Different Post-acquisition Feeding Times.
Identification of Endogenous Genes for Normalizing Titer Variation of Citrus Tristeza Virus in Aphids at Different Post-acquisition Feeding Times.
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DOI：
10.5423/ppj.oa.01.2022.0011
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发表时间：
2022-08
影响因子：
2.3
通讯作者：
Liu, Jinxiang
中科院分区：
文献类型：
--
作者：
Wang, Hongsu;Chen, Qi;Liu, Luqin;Zhou, Yan;Wang, Huanhuan;Li, Zhongan;Liu, Jinxiang
柑橘衰退病毒（Citrus tristeza virus，CTV）是柑橘褐蚜（Toxoptera citricida（Kirkaldy））以半持续方式传播的一种病毒。目前，用于检测昆虫载体中的植物病毒的最灵敏的方法是逆转录定量聚合酶链反应（RT-qPCR）。在本研究中，延伸因子-1 α（EF-1α）基因和酸性p0核糖体蛋白（RPAP 0）基因被证实是合适的参考基因，用于RT-qPCR标准化在含病毒T. citricida蚜虫使用geNorm，Normalware，和BestKeeper工具.然后测定了蚜虫体内CTV的相对滴度（T.使用EF-1α和RPAP 0作为参考基因，通过RT-qPCR对在健康植物上不同采集后饲喂时间的柠檬酸单胞菌（citricida）进行定量。随着取食时间的延长，蚜虫体内保留的CTV相对滴度逐渐降低。在健康植物上的第一个0.5小时的喂养时间期间，蚜虫中剩余的CTV滴度显示出对于高传播性分离物CT 11 A约80%的快速损失和对于低传播性分离物CTLJ约40%的损失。对于CT 11 A，在采集后超过12小时期间蚜虫中的相对CTV滴度显著低于其他饲喂时间，这与CTLJ发现的趋势相似。据我们所知，这是第一次报道CTV在T. citricida在不同的收购后的健康植物上的喂养时间。
Citrus tristeza virus (CTV) is efficiently transmitted in a semi-persistent manner by the brown citrus aphid (Toxoptera citricida (Kirkaldy)). Currently, the most sensitive method for detecting plant viruses in insect vectors is reverse-transcription quantitative polymerase chain reaction (RT-qPCR). In this study, the elongation factor-1 alpha (EF-1α) gene and acidic p0 ribosomal protein (RPAP0) gene were confirmed to be suitable reference genes for RT-qPCR normalization in viruliferous T. citricida aphids using the geNorm, NormFinder, and BestKeeper tools. Then the relative CTV titer in aphids (T. citricida) at different post-acquisition feeding times on healthy plants was quantified by RT-qPCR using EF-1α and RPAP0 as reference genes. The relative CTV titer retained in the aphids gradually decreased with increasing feeding time. During the first 0.5 h of feeding time on healthy plants, the remaining CTV titer in aphids showed about 80% rapid loss for the highly transmissible isolate CT11A and 40% loss for the poorly transmissible isolate CTLJ. The relative CTV titer in aphids during more than 12 h post-acquisition times for CT11A was significantly lower than at the other feeding times, which is similar to the trend found for CTLJ. To our knowledge, this is the first report about the relative titer variation of CTV remaining in T. citricida at different post-acquisition feeding times on healthy plants.